Characterization of the inflammatory infiltrate and cytokine expression in the skin of horses with recurrent urticaria

Background – Recurrent urticaria (RU) is a common skin disease of horses, but little is known about its pathogenesis. Hypothesis/Objective – The aim of this study was to characterize the inflammatory cell infiltrate and cytokine expression pattern in the skin of horses with RU. Animals – Biopsies of lesional and nonlesional skin of horses with RU (n = 8) and of skin from healthy control horses (n = 8) were evaluated. Methods – The inflammatory cell infiltrate was analysed by routine histology. Immunohistochemistry was used to identify T cells (CD3), B cells (CD79), macrophages (MAC387) and mast cells (tryptase). Expression of T‐helper 2 cytokines (interleukins IL‐4, IL‐5 and IL‐13), a T‐helper 1 cytokine (interferon‐γ), IL‐4 receptor α and thymic stromal lymphopoietin was assessed by quantitative RT‐PCR. Results – In subepidermal lesional skin of RU‐affected horses, increased numbers of eosinophils (P ≤ 0.01), CD79‐positive (P ≤ 0.01), MAC387‐positive (P ≤ 0.01) and tryptase‐positive cells (P ≤ 0.05) were found compared with healthy horses. Subepidermal lesional skin of RU‐affected horses contained more eosinophils (P ≤ 0.05) and tryptase‐positive cells (P ≤ 0.05) compared with nonlesional skin. There was no significant difference in infiltrating cells between nonlesional skin and skin of healthy horses. Expression of IL‐4 (P ≤ 0.01), IL‐13 (P ≤ 0.05), thymic stromal lymphopoietin (P ≤ 0.05) and IL‐4 receptor α (P ≤ 0.05) was increased in lesional skin of RU‐affected horses compared with control horses. Expression of IL‐4 was higher (P ≤ 0.05) in lesional compared with nonlesional RU skin. Conclusions and clinical importance – Analysis of cytokine expression and inflammatory infiltrate suggests that T‐helper 2 cytokines, eosinophils, mast cells and presumptive macrophages play a role in the pathogenesis of equine RU.     

Specific Polymerase Chain Reaction Identification of Venturia nashicola Using Internally Transcribed Spacer Region in the Ribosomal DNA

ABSTRACT A technique based on the polymerase chain reaction (PCR) was developed for the identification of Venturia nashicola using nucleotide sequence information of the ribosomal DNA region. The complete internal transcribed spacer (ITS) region of V. nashicola strains and phylo-genetically related species was amplified with the two universal ITS1 and ITS4 primers, sequenced, and digested with five restriction enzymes. The alignment of nucleotide sequences and analyses of digestion patterns indicated constant polymorphisms between V. nashicola and related species at nucleotides 126 and 127, which overlapped a TaqI restriction site. An oligonucleotide primer named A126 was designed for identifying this variable region. A primer set (A126 and ITS4) that allowed the amplification of a 391-bp DNA fragment within the ITS region by PCR was specific to V. nashicola when it was checked against fungal genomic DNAs of related fungi. This primer set was a good candidate for a species-specific reagent in a procedure for identification of V. nashicola by PCR.     

Electrophoretic protein profiles of strains of Ceratocystis fimbriata f. sp. platani

Soluble proteins extracted from the mycelium of strains of Ceratocystis fimbriata (Ell. et Halst) Davidson f. sp. platani Walter were separated on native and SDS-polyacrylamide gradient gel electrophoresis. The protein patterns were compared and the electrophoretic profiles of all the strains examined showed striking intraspecific similarities. The use or protein banding patterns as a diagnostic criterion to distinguish strains of C. fimbriata is discussed.     

Sporotrichosis: the main differential diagnosis with tegumentary leishmaniosis in dogs from Rio de Janeiro, Brazil

Seventy-four dogs from the State of Rio de Janeiro with ulcerated cutaneous lesions were submitted to clinical, dermatological, parasitological, mycological, histopathological and cytopathological exams, a leishmanin skin test, an indirect immunofluorescence (IIF) test for leishmaniosis, and nonspecific laboratory tests such as blood count and serum biochemistry. Sporothrix schenckii was isolated from 41 dogs and Leishmania (Viannia) braziliensis was isolated from 33 animals. Most dogs with sporotrichosis were from the municipality of Rio de Janeiro (53.7%) and presented ulcerated cutaneous lesions on the head (68.3%). Laboratory alterations in these animals included anemia (58.5%), hypoalbuminemia (83%) and hyperglobulinemia (75.6%). Histopathology revealed the predominance of a chronic granulomatous inflammatory infiltrate (70.7%), and yeast-like structures were detected in 17% of the histopathological exams and in 32% of the cytological exams. Three of 41 dogs with sporotrichosis were seropositive by IIF for leishmaniosis and 2 of 20 animals tested within this group had a positive leishmanin skin test. Similarly, most of the 33 dogs with leishmaniosis were from the municipality of Rio de Janeiro (69.7%) and had ulcerated cutaneous lesions on the head (84.8%). Laboratory alterations in these animals included anemia (66.7%), hypoalbuminemia (100%) and hyperglobulinemia (91%). Histopathology showed the predominance of a chronic granulomatous inflammatory infiltrate (63.6%) and amastigote forms were detected in 30.3% of the histopathological exams and in 31.8% of the 22 cytological exams performed. About 72.7% of the dogs were seropositive by IIF and five of seven animals had a positive skin test. Due to the clinical similarities, histopathological and nonspecific laboratory results similarities, the serological and skin tests for leishmaniosis positive in dogs with sporotrichosis, and the overlapping endemic areas in Rio de Janeiro, the differential diagnosis between the two diseases requires the demonstration of their respective etiological agents.     

A selective and low impacting traditional fishery,sustaining the economy of small coastal villages in central Mediterranean: Keep or replace the small‐scale driftnets?

Small‐scale driftnets (SSDs) have been historically used in the Mediterranean without major environmental concern. The introduction of large‐scale driftnets caused unwanted catches of protected species. Specific regulations were therefore issued in European waters, culminating in a proposed moratorium on SSDs. This study aimed to characterise the SSD fishery targeting anchovy (menaide), evaluating its environmental sustainability, economic performance and social relevance. In 2013, a survey by interviews, logbooks and observers on board assessed the order of magnitude of this fishery in terms of fishing capacity and activity, volume of landings and revenues. The menaide fleet consisted in 60 vessels <12 m overall length, moored in little harbours in southern Italy. These nets are highly selective: the target species, European anchovy, Engraulis encrasicholus (L.) and sardine, Sardina pilchardus (Walbaum), dominates the catches, while discards are negligible. The anchovies caught are of high quality and large size; the high prices support fish processing by local factories. Moreover, the results of a SWOT analysis demonstrated that replacing SSDs with a semi‐industrial fishery, like purse seining, would increase impacts on the ecosystem, and a loss of socio‐economic opportunities for several coastal villages. These findings support the option of keeping SSDs operating, in the framework of specifically oriented management measures.     

Antimicrobial activity of Tunisian quince (Cydonia oblonga Miller) pulp and peel polyphenolic extracts

Quince (Cydonia oblonga Miller) fruit aqueous acetone extracts were evaluated. High-performance liquid chromatography-diode array detection and electrospray ionization-mass spectrometry were used for the identification and quantification of the phenolic compounds. The total phenolic content of the pulp and peel parts ranged from 37 to 47 and 105 to 157 mg/100 g of fresh weight, respectively. Chlorogenic acid (5-O-caffeoylquinic acid) was the most abundant phenolic compound in the pulp (37%), whereas rutin (quercetin 3-O-rutinoside) was the main one in the peel (36%). The radical scavenging potential of the extracts was determined and compared with that of synthetic antioxidants. The stronger properties corresponded to those obtained from peel material with a 70-80% inhibitory effect on DPPH radicals. The antimicrobial activity of the extracts against different microorganism strains was also investigated. Quince peel extract was the most active for inhibiting bacteria growth with minimum inhibitory and bactericide concentrations in the range of 102-5 x 103 microg polyphenol/mL. It seems that chlorogenic acid acts in synergism with other components of the extracts to exhibit their total antimicrobial activities.     

Gully erosion modelling and landscape response in the Mbuluzi River catchment of Swaziland

In southern African countries soil erosion and the related problems, such as water quality issues or decreasing soil productivity, are the main topics affecting the inhabitants of both rural and urban areas. Therefore, the attention has been recently placed on those problems related to soil erosion. This can also be documented by an increasing number of studies carried out on erosion and by the development and application of erosion models. Nevertheless, gully erosion phenomena have been widely neglected in erosion modelling. This is because the development of erosion models was focused on those regions with an intense agriculture typical of developed countries on the one hand, and because of the spatial and temporal heterogeneity of gully erosion processes on the other hand. This study regards the identification of gully erosion forms and processes in the Mbuluzi River catchment (Kingdom of Swaziland) by using the Erosion Response Units (ERU) concept. The following modelling of gully erosion was done through the stable gully model [Catena 37 (1999) 401]. The input data were obtained through the application of remote sensing techniques (API method) and GIS-analyses. The example from Swaziland shows that the applied methods are able to identify areas affected by gully erosion. Furthermore, it is possible to estimate the amount of soil loss due to gully erosion, which, for example, is not taken into consideration by the USLE-type models.     

Ultrasensitive quantitative determination of paraquat: application to river, ground, and drinking water analysis in an agricultural area

The water specimens were collected from wells and irrigation ditches in the agricultural area to the south of Milan and from Olona River and Mantua Lake and analyzed for paraquat detection. The assay was performed using a specific polyclonal antibody raised in sheep and rabbit anti-sheep IgG conjugated with a chelating molecule 4,7-bis(chlorosulfophenyl)-1,10-phenanthroline-2,9-dicarboxylic acid complexed with Eu3+ as a fluorescent marker. Bovine serum albumin conjugated with 5-(1'-methyl[4,4']bipyridinyl-1-yl)pentanoic acid was used in solid phase preparation. The sensitivity achieved was 20 ng L(-1). The recovery in samples spiked with three different PQ concentrations was between 88 and 108%.     

Characterizing the molecular and morphophysiological diversity of Italian red clover

Red clover (Trifolium pratense L.) is the third major forage species in Europe, but there is limited information on the biodiversity and the genetic structure of landraces and natural populations which evolved in this region. The objective of this study was producing such information for Italian germplasm on the ground of molecular and morphophysiological diversity. The study included 16 Italian natural populations from a wide range of environments, four landraces representing the four traditional commercial ecotypes, and two varieties. Eight morphophysiological traits were assessed in a replicated trial under field conditions, whereas two AFLPs primer combinations with 140 polymorphic markers were recorded on a random sample of 13 genotypes per population. Ordination and classification results based on morphophysiological data clearly kept track of the type of germplasm (i.e. landrace or natural population) and its geographic origin, unlike results based on molecular markers. Euclidean distances among populations based on morphophysiological traits were not correlated with Nei’s genetic distances based on molecular markers according to Mantel’s test. Geographical distances among landrace or natural population material was correlated with distances based on morphophysiological traits but not with those based on molecular markers. The average within-population variation estimated via molecular markers was about 2.6-fold higher than that among populations, preventing an acceptable discrimination among most populations. On average, natural populations tended to have within-population variation similar to varieties and somewhat lower than landraces. Our results have implications for collection, conservation, exploitation and registration in a sui generis system of red clover genetic resources.     

Metabolites from Marine Sponges and Their Potential to Treat Malarial Protozoan Parasites Infection: A Systematic Review

Malaria is an infectious disease caused by protozoan parasites of the Plasmodium genus through the bite of female Anopheles mosquitoes, affecting 228 million people and causing 415 thousand deaths in 2018. Artemisinin-based combination therapies (ACTs) are the most recommended treatment for malaria; however, the emergence of multidrug resistance has unfortunately limited their effects and challenged the field. In this context, the ocean and its rich biodiversity have emerged as a very promising resource of bioactive compounds and secondary metabolites from different marine organisms. This systematic review of the literature focuses on the advances achieved in the search for new antimalarials from marine sponges, which are ancient organisms that developed defense mechanisms in a hostile environment. The principal inclusion criterion for analysis was articles with compounds with IC₅₀ below 10 µM or 10 µg/mL against P. falciparum culture. The secondary metabolites identified include alkaloids, terpenoids, polyketides endoperoxides and glycosphingolipids. The structural features of active compounds selected in this review may be an interesting scaffold to inspire synthetic development of new antimalarials for selectively targeting parasite cell metabolism.